CHOgro Transfection and Titer Enhancer Kit

Description

A Superior Transient Expression System for CHO Cells with the CHOgro Titer Enhancer

The CHOgro® High Yield Expression System Features CHOgro® Titer Enhancer. The CHOgro® High Yield Expression System (MIR 6270) includes the same components of the first-generation CHOgro® Expression System (MIR 6260) and features a new component – the CHOgro® Titer Enhancer [which is also included in the CHOgro® Transfection and Titer Enhancer Kit (MIR 6225)]. Adding the CHOgro® Titer Enhancer and shifting the cell culture temperature to 32°C yields a significantly higher increase in in titers over the original CHOgro® Expression System.

Kit includes: TransIT-PRO® Transfection Reagent (1 ml), CHOgro® Titer Enhancer (20 ml)

SKU: MIR 6225

Supporting Data

CHOgro System-to-System Comparison

CHOgro® High Yield Expression System Streamlined Workflow. The CHOgro® High Yield Expression System was engineered to maximize transient protein production in suspension CHO cells, while still maintaining a simple and cost-effective workflow. The ability to add expression enhancers at the time of transfection and immediately shift cell cultures to hypothermic conditions provides researchers with more flexibility in the timing of their experiments while reducing the risk of contamination caused by repeated handling of the culture.

CHOgro® High Yield Expression System Results in Higher Protein Titers. The CHOgro® High Yield Expression System (MIR 6270) and the CHOgro® Expression System (MIR 6260) were compared for human IgG1 antibody production. Antibody was produced by transient transfection using the TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt) and 1 µg plasmid DNA per milliliter of culture. FreeStyle™ CHO-S cells (ThermoFisher Scientific) were cultured in CHOgro® Expression Medium (Mirus Bio) and plated at a density of 2 or 4 x 10⁶ cells/ml in non-treated 6-well plates (2ml/well) for transfection. As indicated, the CHOgro® High Yield culture was shifted to 32°C immediately post-addition of the transfection complexes to the culture. Day 10 supernatants were analyzed using a standard sandwich human IgG ELISA. Error bars represent the standard deviation of triplicate technical replicates.

CHOgro® Titer Enhancer Does Not Adversely Affect Cell Growth and Viability Post-transfection. Triplicate flasks of FreeStyle™ CHO-S cells adapted to CHOgro® Expression Medium were transiently transfected with the TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt) and 1 µg plasmid DNA per milliliter of culture at 4 x 10⁶ cells/ml in 125 ml Optimum Growth flasks (25 ml/flask, Thomson Instrument Company). As indicated, CHOgro® Titer Enhancer was added and all of the cultures were shifted to 32°C immediately post-addition of the transfection complexes to the culture. Cell counts (solid line) and viability (propidium iodide staining, dotted line) were measured daily post-transfection using a Guava® easyCyte™ 5HT flow cytometer (EMD Millipore).

TransIT-PRO® + CHOgro® Titer Enhancer Outperforms PEImax and linear 25 kDa PEI Transfection Reagents. An IgG1 antibody construct was produced by transient transfection using either the TransIT-PRO® Transfection Reagent (Mirus Bio) at a 1:1 reagent-to-DNA ratio (vol:wt), PEImax (4:1, Polysciences), or linear 25 kDa PEI (6:1, Polysciences) and 1 µg plasmid DNA per milliliter of culture in FreeStyle™ CHO-S cells (ThermoFisher Scientific) cultured in CHOgro® Expression Medium (Mirus Bio) at a cell density of 4 x 10⁶ cells/ml. CHOgro® Titer Enhancer (Mirus Bio) was added immediately post-addition of transfection complexes and cultures were incubated at 32°C with shaking until harvest for all reagents. Day 14 supernatants were analyzed using a standard sandwich human IgG ELISA. Error bars represent the standard deviation of triplicate technical replicates.

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Specifications

Storage Conditions
CHOgro® High Yield Expression System: Multiple storage conditions – see individual bottles for specific recommendations
CHOgro® Transfection and Titer Enhancer Kit: Multiple storage conditions – see individual bottles for specific recommendations
CHOgro® Expression Medium: Store at 4°C
TransIT-PRO® Transfection Reagent: Store at -20°C
CHOgro® Complex Formation Solution: Store at 4°C
Poloxamer 188 Solution, 10%: Store at room temperature
L-Glutamine Solution, 200 mM: Store at -20°C

Product Guarantee
CHOgro® High Yield Expression System: As labeled on the product components
CHOgro® Transfection and Titer Enhancer Kit: As labeled on the product components
CHOgro® Expression Medium: As labeled on the product
TransIT-PRO® Transfection Reagent: 1 year
CHOgro® Complex Formation Solution: As labeled on the product
Poloxamer 188 Solution, 10%: As labeled on the product
L-Glutamine Solution, 200 mM: As labeled on the product

Usage Statement
All Configurations: For Research Use Only.

Animal Origin Statement
All Configurations: This product is animal origin free.